Analyzing Satellite Cell Function During Skeletal Muscle Regeneration by Cardiotoxin Injury and Injection of Self-delivering siRNA In Vivo
Analyzing Satellite Cell Function During Skeletal Muscle Regeneration by Cardiotoxin Injury and Injection of Self-delivering siRNA In Vivo
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DOI：
10.3791/60194
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发表时间：
2019-09-01
影响因子：
1.2
通讯作者：
von Maltzahn, Julia
中科院分区：
文献类型：
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作者：
Ahrens, Hellen E.;Henze, Henriette;von Maltzahn, Julia
骨骼肌在受伤后具有巨大的再生能力。这一过程主要由肌肉干细胞驱动，也称为卫星细胞。卫星细胞的特征是转录因子Pax7的表达，以及它们位于静息骨骼肌基板下的位置。损伤后，卫星细胞被激活，经历自我更新或分化，形成新的肌纤维或与受损的肌纤维融合。利用基于心脏毒素的骨骼肌损伤模型，可以研究体内卫星细胞的功能。为了研究单个基因在骨骼肌再生过程中的作用，通常采用转基因小鼠模型。在这里，我们提出了一种替代转基因小鼠的方法，来研究再生过程中卫星细胞中的基因功能，例如，在没有转基因小鼠的情况下。我们将心脏毒素介导的特定骨骼肌损伤与将自我递送的siRNA注射到再生肌肉中，然后由卫星细胞和其他细胞吸收。因此，我们提供了一种在生理条件下分析卫星细胞再生过程中基因功能的方法，而不需要转基因小鼠。
Skeletal muscle possesses an enormous capacity to regenerate after injury. This process is mainly driven by muscle stem cells, also termed satellite cells. Satellite cells are characterized by the expression of the transcription factor Pax7 and their location underneath the basal lamina in the resting skeletal muscle. Upon injury, satellite cells get activated, undergo self-renewal or differentiation to either form new myofibers or to fuse with damaged ones. The functionality of satellite cells in vivo can be investigated using a cardiotoxin based injury model of skeletal muscle. To study the function of one gene during the regeneration of skeletal muscle, transgenic mouse models are mostly used. Here, we present an alternative method to transgenic mice, to investigate the gene function in satellite cells during regeneration, e.g., in cases where transgenic mice are not available. We combine the cardiotoxin mediated injury of a specific skeletal muscle with the injection of a self-delivering siRNA into the regenerating muscle which is then taken up by satellite cells among other cells. Thereby, we provide a method to analyze gene function in satellite cells during regeneration under physiological conditions without the need for transgenic mice.